Figure 8.

Purinosome Components Regulate Cortical Development

(A–R) A control shRNA or Paics shRNA (shRNA #1 or shRNA #3) was delivered into the brain on E14.5, together with EGFP, and the cortices were analyzed at E18.5. (A and B) Distribution of EGFP⁺ cells in the neocortex. (C) Quantification of the distribution of EGFP⁺ cells in the indicated areas. ∗∗p < 0.01. (D–R) Brain sections of Paics knockdown were immunostained using anti-Dcx (D–F), anti-Pax6 (J–L), anti-Tbr2 (G–I), and anti-Ki67 (M–O) antibodies. The VZ surface is outlined by the dashed line. (P–R) Higher magnification of the boxed area depicted in (O). Many NSPCs electroporated with the Paics shRNAs remained in the SVZ/VZ/IZ as postmitotic cells that were Dcx⁺ Pax6^– Tbr2^–.

(S–Y) Control EGFP or Fgams-EGFP was delivered into the brain on E14.5, and the cortices were analyzed at E16.5 (S and T) or E18.5 (V, W, and Y). (U and X) Distribution of EGFP⁺ cells in the indicated areas at E16.5 (U) or E18.5 (X). ∗p < 0.05, ∗∗∗p < 0.001. (Y) Fgams-EGFP-overexpressing brain immunostained with an anti-Nestin antibody. Forced expression of Fgams-EGFP directed many NSPCs to remain in the SVZ/VZ as Nestin⁺ cells, and only a few cells reached the border between the SVZ and IZ. All data are presented as means ± SEM. Statistical significance value was determined using the Welch's t test followed by Holm-Bonferroni correction.

Scale bars, 100 μm in (A)–(O) and (S)–(Y); 15 μm in (P)–(R).